• Title/Summary/Keyword: Co-incubated

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Effect of a short-term in vitro exposure time on the production of in vitro produced piglets

  • Hwang, In-Sul;Kwon, Dae-Jin;Kwak, Tae-Uk;Lee, Joo-Young;Hyung, Nam-Woong;Yang, Hyeon;Oh, Keon Bong;Ock, Sun-A;Park, Eung-Woo;Im, Gi-Sun;Hwang, Seongsoo
    • Journal of Embryo Transfer
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    • v.31 no.2
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    • pp.117-121
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    • 2016
  • Although piglets have been delivered by embryo transfer (ET) with in vitro produced (IVP) embryos and blastocysts, a success rate has still remained lower level. Unlike mouse, human, and bovine, it is difficult to a production of piglets by in vitro fertilization (IVF) because of an inappropriate in vitro culture (IVC) system in pig. Therefore, the present study was conducted to investigate whether minimized exposure time in IVC can improve the pregnancy and delivery rates of piglets. Immediately after IVM, the oocytes were denuded and co-incubated with freshly ejaculated boar semen for 3.5 to 4 hours at $38.5^{\circ}C$ under 5% $CO_2$ in air. To avoid long-term exposure to in vitro state, we emitted IVC step after IVF. After that the presumptive zygotes were transferred into both oviducts of the surrogate on the same day or 1 day after the onset of estrus. Pregnancy was diagnosed on day 28 after ET and then was checked regularly every month by ultrasound examination. The 3 out of 4 surrogates were determined as pregnant (75%) and a total of 5 piglets (2 females and 3 males) were delivered at $118.3{\pm}2.5$ days of pregnancy period. In conclusion, a short-term exposure time may be an important factor in the production of IVP-derived piglets. It can be apply to the in vitro production system of transgenic pig by IVF, cloning, and pronuclear microinjection methods.

Formation of D-Glucose Isomerase by Streptomyces sp. (Streptomyces sp.에 의한 포도당 이성화효소의 생성)

  • Rhee, In-Koo;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.8 no.3
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    • pp.173-180
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    • 1980
  • A source of D-xylose was required for the enhanced production of D-glucose isomerase of Streptomyces sp. strain K-17. D-glucose supported the luxuriant growth of the organism as well as D-xylose, but D-glucose isomerase activity was hardly detected in the D-glucose-grown cells. When the D-glucose-grown cells were incubated aerobically for a few hours in 0.5% xylose solution in 0.05 M phosphate buffer, pH 7.0, it was found that inductive formation of D-glucose isomerase occurred in the cells without multiplication. In the non-growth phase of cells the inductive formation of D-glucose isomerase occurred because a source of nitrogen for the synthesis of enzymes was obtained from turnover of protein accumulated in cells. D-ribose, L-arabinose, D-glucose, D-mannose, citrate, succinate and tartrate could not induce the formation of D-glucose isomerase, but D-xylose could induce. Inductinn of D-glucose isomerase was repressed by D-glucose and its catabolites : glycerol, succinate and citrate. Inductive formation of the enzymes in the non-growth phase was stimulated by $Ba^{2+}$, $Mg^{2+}$ and $Co^{2+}$, and inhibited by C $u^{2+}$, C $d^{2+}$, A $g^{+}$and H $g^{2+}$. The synthesis of enzymes in the induction system composed of 0.5% xylose solution was disrupted by actinomycin D, streptomycin, chloramphenicol, kanamycin, tetracycline, p-chloromercuribenzo ate, arsenate and 2, 4-dinitrophenol, but not disrupted by mitomycin C and penicillin G.icillin G.

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Feasibility of Analyzing Soil Organic Carbon Fractions using Mid-Infrared Spectroscopy (중적외선분광분석법을 이용한 토양 유기 탄소 분획 분석)

  • Hong, Seung-Gil;Shin, JoungDu;Park, Kwang-Lai;Lee, Sang-Beom;Kim, Jinho;Kim, Seok-Cheol;Shiedung, Henning;Amelung, Wulf
    • Journal of the Korea Organic Resources Recycling Association
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    • v.23 no.3
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    • pp.85-92
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    • 2015
  • For concerning the climate change issues, the carbon sequestration and importance of soil organic matter are receiving high attention. To evaluate carbon sequestration in soil is important to determine the soil organic carbon (SOC) fractions such as WESOC (Water extractable soil organic carbon), and $CO_2$ emission by soil microbial respiration. However, the analyses for those contents are time-consuming procedure. There were studied the feasibility of MIRS (Mid-Infrared Spectroscopy), which has short analysis time for determining the WESOC and an incubated carbon in this study. Oven-dried soils at $100^{\circ}C$ and $350^{\circ}C$ were scanned with MIRS and compared with the chemically analyzed WESOC and cumulative carbon dioxide generated during 30, 60, 90, and 120 days of incubation periods, respectively. It was observed that an optimized determination coefficient was 0.6937 between WESOC and untreated soil processed by spectrum vector normalization (SNV) and 0.8933 between cumulative $CO_2$ from 30 days incubation and soil dried at $350^{\circ}C$ after subtracting air-dried soil processed by 1st derivatives. Therefore, it was shown that Quantification of soil organic carbon fractions was possibility to be analyzed by using MIRS.

Evaluation of Cytotoxic Properties of Tea Polyphenols in Intestinal Cells Treated with Over-the-counter Drugs (녹차 폴리페놀 성분과 일반 의약품의 상호작용에 의한 장관계 세포 독성 평가)

  • Choi, Hyun-A;Kim, Mi-Ri;Hong, Jung-Il
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.641-647
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    • 2011
  • Polyphenols in green tea are biologically active and may interact with commonly-consumed over-the-counter (OTC) drugs in the body. In this study, modulation of cytotoxicity of polyphenon 60 (PPE, tea polyphenol mixture) with co-treatment of several OTC drugs, including ibuprofen (Ibu), acetaminophen (AAP), and aspirin was investigated in intestinal cells. PPE showed more potent cytotoxic effects on colon cancer HCT 116 cells than on normal intestinal INT 407 cells. Ibu had the strongest cytotoxic effects on both cell types. Cytotoxicity of PPE on HCT 116 and INT 407 cells was not markedly altered by co-treated OTC drugs. Cytotoxicity of the OTC drugs was not affected by PPE. When HCT 116 cells were incubated with AAP before or after PPE treatment, cytotoxicity was slightly enhanced more than their additive effect. The present study may provide basic information of possible toxicity due to interaction of the polyphenols and the OTC drugs.

Transformation of Pentachlorophenol by Oxidoreductive Catalysts (산화환원 촉매에 의한 Pentachlorophenol의 Transformation)

  • Park, Jong-Woo;Lee, Yun-Ki;Kim, Jang-Eok
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.330-335
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    • 1999
  • Pentachlorophenol(PCP), which is very persistent in soil and water environment, was tried to detoxify with oxidoreductive catalysts(peroxidase, laccase, tyrosinase and birnessite). To find out detoxification of PCP, the transformation of PCP through oxidative coupling was investigated in the presence of various oxidoreductive catalysts. PCP incubated with peroxidase was significantly transformed, however, in case of tyrosinase, the transformation was negligible. Using peroxidase, the optimal reaction condition was pH 5.6 and $16^{\circ}C$. The transformation of PCP was very fast in initiation step until 30 min but, that was not observed after 180 min. The transformation of PCP was increased by increasing peroacidase amount. When the effect of humic monomer was investigated as co-substrate on the transformation of PCP, the transformation of PCP was mostly decreased in the incubation with peroxidase, laccase, and birnessite. The transformation of PCP, however, was slightly increased by the incubation with tyrosinase in the presence of humic monomers as co-substrate, except catechol. On the basis of the results obtained, it may be suggested that PCP is able to be effectively detoxified through oxidative coupling mediated with oxidoreductive catalysts.

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Changes in the CO2 and amount of mycelium growth of the liquid spawn on Flammulina velutipes (팽나무버섯 액체 종균 배양시 이산화탄소 농도와 균사 생장량 변화)

  • Shim, Kyu-Kwang;Yoo, Young-Jin;Koo, Chang-Duck;Kim, Young-Seok;Kim, Myung-Koon
    • Journal of Mushroom
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    • v.10 no.1
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    • pp.3-8
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    • 2012
  • In this study, to produce Flammulina velutipes mushroom liquid spawn efficiently and effectively the effects of explosive aeration (supplying air with tiny bubbles) of the liquid culture medium on carbon dioxide concentration and residual sugar content in the medium on carbon dioxide concentration and residual sugar contentin the medium were measured. Carbon dioxide concentrations were measured at the outlet of the incubator. On the third day the explosive aeration greatly increased mycelial growth of the liquid spawn, and carbon dioxide concentration also greatly increased but decreased after 5 days. Free sugar contents in the liquid culture consistantly decreased up to 7 days and thereafter was not detected. The weight of the mycelia were maintained similar levels after 3 days. Total nitrogen content in the liquid medium constantly decreased during the 11days of explosive aeration. The content of free sugars in 7 days of culture was the lowest level, thus the inoculum incubated for 6~7 days was thought to be the most effective. Carbon dioxide concentration measurement at the outlet of the container during the liquid spawn incubation required low cost but was efficient to estimate the degree of mycelial growth to be used as a simple indicator.

The Cytotoxic Effect of Vibrio vulnificus Hemolysin on the Mouse Peritoneal Macrophages (마우스 복강내 대식세포에 대한 Vibrio vulnificus Hemolysin의 세포독성)

  • Im, Ihn-Soo;Lee, Shee-Eun;Kim, Seol;Bae, Mi-Ok;Rhee, Joon-Haeng;Shin, Boo-Ahn;Cung, Sun-Sik;Ryu, Phil-Youl
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.251-261
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    • 2000
  • V. vulnificus is an estuarine bacterium which causes septicemia and shock in susceptible patients. The organism produces a hemolytic cytolysin (VvH), which has a membrane damaging effect on erythrocytes. To clarify the mechanisms by which VvH might contribute to virulence, we examined its effect on macrophages. When mouse peritoneal macrophages were harvested and co-cultured with hemolysin-positive V. vulnificus strains (100 bacteria/cell), about 60% of the macrophages were killed; macrophages were not killed when co-cultured V. vulnificus strain CVD 707, a VvH-negative deletion mutant. Exposure of macrophages to filtered culture supernatants (2.5 HU/ml) and purified VvH (3 HU/ml) resulted in an increase in dead cells (80 and 90%, respectively), as determined by the trypan blue dye exclusion method and LDH release from macrophages was also increased (70 and 65.5%, respectively). The cytotoxic effect of VvH on macrophages was both the dose- and time-dependent. The VvH caused damage to the macrophage membrane and was blocked significantly by preincubation with cholesterol (p<0.01). Fetal bovine serum showed remarkable inhibition of VvH synthesis by V. vulnificus and inhibited VvH activity in culture supernatant. Cell viability was increased by 35% (p<0.01) and LDH release decreased by 28% (p<0.01) when macrophages were incubated with V. vulnificus (100 bacterial cell) in DMEM-10% FBS for 2 hr. Bacterial clearance activity of mice against V. vulnificus CVD 707 was decreased by pretreatment with 10 HU of VvH. This result suggests that the VvH can impair the membrane of macrophages and may playa role in the pathogenesis of V. vulnificus septicemia.

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Eleutherosides Extraction from Acanthopanax sessiliflorus Seeman and Eleutherococcus senticosus Maxim Using an Enzymatic Process (효소적 생물전환을 이용한 토종오가피와 가시오가피의 Eleutheroside B, E의 추출 함량 최적화)

  • Kim, Na-Ri;Park, Jong-Soon;Lee, Deuk-Sik;Shim, Jae-Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.9
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    • pp.1273-1278
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    • 2016
  • In this study, we optimized conditions for extraction of eleutherosides B and E from stem powder of Acanthopanax. To enhance eleutheroside B and E yields, Acanthopanax sessiliflorus Seeman and Eleutherococcus senticosus Maxim were pre-incubated with the following enzymes: Celluclast, Viscozyme L, Lactozyme, Lecitase, and Novozyme 33095. Treatment with Novozyme 33095, a commercial pectinase, for 3 h resulted in the highest yields of eleutherosides B and E from A. sessiliflorus and E. senticosus. Compared with extraction at $121^{\circ}C$ for 120 min, at $121^{\circ}C$ for 15 min after Novozyme 33095 pre-incubation increased eleutheroside B and E yields from A. sessiliflorus by 7% and 17%, respectively. In the case of E. senticosus, eleutheroside B and E yields increased by 25% and 29%, respectively.

Effects of Sodium Nitroprusside and Nitric Oxide on In Vitro Maturation of Canine Oocytes

  • Park, Ji-Hoon;Lee, Bong-Koo;Chung, Young-Ho;Rhee, Man-Hee;Kim, S.K.
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.469-473
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    • 2011
  • These study was carried out to investigate the effects of the supplementation with sodium nitroprusside (SN) and nitric oxide (NO) of canine oocytes on IVM rates. Oocytes were incubated in TCM-199 supplement with at 0.03~0.10 mM SN and 0.3~1.0 mM NO for 48 hrs. Oocytes were transferred to 50 ul drops of maturation medium covered mineral oil and cultured in a $CO_2$ incubator (5% $CO_2$, 95% air, $38^{\circ}C$). The in vitro maturation rate of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.03, 0.05, 0.07, 0.10 mM SN were $25.9{\pm}3.5%$, $36.4{\pm}3.2%$, $33.3{\pm}3.5%$, $28.8{\m}3.2%$, respectively. The in vitro maturation rate of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.03~0.07 mM SN were significantly increased compare to the control ($26.0{\pm}2.2%$). The in vitro maturation rates of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.3, 0.5, 0.7, 1.0 mM NO were $28.0{\pm}4.2%$, $36.5{\pm}3.6%$, $30.0{\pm}3.8%$, $19.2{\pm}3.5%$, respectively. The in vitro maturation rate of oocytes in TCM-199 medium supplemented with 0.3 and 0.5 mM NO were significantly increased compare to the control ($26.0{\pm}2.2%$). The in vitro maturation rates of oocytes cultured for 12~48 hrs in TCM-199 medium supplement with 0.05 mM SN were $26.0{\pm}3.2%$, $28.0{\pm}3.4%$, $38.0{\pm}3.2%$, respectively. The in vitro maturation rate of oocytes cultured for 12~48 hrs in TCM-199 medium supplement with 0.5 mM NO were $22.0{\pm}3.0%$, $30.0{\pm}3.8%$, $36.0{\pm}4.2%$, respectively. These result was significantly increased compare to the control.

THE EFFECT OF THE SAPONIN FRACTION OF PANAX GINSENG C.A. MEYER ON THE ANTIOXIDANT ACTIVITY OF TOCOPHEROL (한국산 인삼(Panax ginseng C. A, Meyer)의 사포닌 성분이 ${\alpha}-tocopherol$의 항산화작용에 미치는 영향)

  • Hong Sa-Duk;Koo Ja-Don;Park Gyeong Suk;Hong Jeong Tae
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.113-118
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    • 1984
  • The effect of the saponin fraction extracted from Panax ginseng C.A. Meyer on the antioxidant activity of ${\alpha}-tocopherol$ was investigated in vitro as well as in vivo. Microsomal preparation of albino rat (Sprague-Dawley, 180-200g) was incubated in the mixture containing NADPH, $Fe^{3+},$ ATP, ${\alpha}-tocopherol$ with and/or without ginseng saponin fraction for 30 minutes and the malondialdehyde formed was assayed and found that the saponin fraction stimulated the antioxidant activity of ${\alpha}-tocopherol$ cooperatively. It was also realized that the cooperative stimulation of the antioxidant activity of ${\alpha}-tocopherol$ was most eminent when the concentration of the saponin fraction was around $10^{-5}\%$ in the reaction mixture. Alcoholic suspension of ${\alpha}-tocopherol$ with and/or without ginseng saponin fraction was administered orally to rats in which the lipid peroxidation was induced by ethanol administration and the lipid peroxide contents of the liver were assayed at certain periods of time after ${\alpha}-tocopherol$ administration in this animal. It was reported that the saponin fraction stimulated the absorption of ${\alpha}-tocopherol$ in rats and this was confirmed again in the present work. From the previous work and present experimental results, it seemed that the saponin fraction accelerated the absorption of ${\alpha}-tocopherol$ and therefore stimulated the antioxidant activity of ${\alpha}-tocopherol$ more effectively in the animal body.

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